Benzothiophenes for treating estrogen deficiency

ABSTRACT

This invention is related to novel benzothiophene compounds of formula I: ##STR1## where R, R 1 , R 2 , X and m are as defined in the specification, which are useful for the inhibition of the various medical conditions associated with postmenopausal syndrome, as well as estrogen dependent diseases including cancer of the breast, uterus, and cervix. The present invention further relates to pharmaceutical formulations of compounds of formula I.

This application claims the benefit of U.S. Provisional Application Ser. No. 60/059,620, filed Sep. 23, 1997.

FIELD OF INVENTION

This invention relates to the fields of pharmaceutical and organic chemistry and provides novel benzothiophene compounds which are useful for the inhibition of the various medical conditions associated with postmenopausal syndrome, as well as estrogen-dependent diseases including cancer of the breast, uterus, and cervix.

BACKGROUND OF THE INVENTION

"Postmenopausal syndrome" is a term used to describe various pathological conditions which frequently affect women who have entered into or completed the physiological metamorphosis known as menopause. Although numerous pathologies are contemplated by the use of this term, three major medical conditions of postmenopausal syndrome are the source of the greatest long-term medical concern: osteoporosis, cardiovascular effects such as hyperlipidemia, and estrogen-dependent cancer such as breast and uterine cancer.

Osteoporosis, which generally includes a group of disorders which arise from diverse etiologies, is characterized by the net loss of bone mass per unit volume. The consequence of this loss of bone mass and resulting bone fracture is the failure of the skeleton to provide adequate structural support for the body. One of the most common types of osteoporosis is that associated with menopause. Most women lose from about 20% to about 60% of the bone mass in the trabecular compartment of the bone within three to six years after the cessation of menses. This rapid loss is generally associated with an increase of bone resorption and formation. However, the resorptive cycle is more dominant and the result is a net loss of bone mass.

Osteoporosis is a common and serious disease among postmenopausal women. There are an estimated 25 million women in the United States who are afflicted with this disease. The results of osteoporosis disease's sequelae are personally harmful and often result in the need for extensive and long term medical support (hospitalization and nursing home care). This is especially true in elderly patients. Additionally, although osteoporosis is not generally thought of as a life threatening condition, a 20% to 30% mortality rate is related with hip fractures in elderly women. A large percentage of this mortality rate can be directly associated with postmenopausal osteoporosis.

The trabecular tissue is the most vulnerable bone tissue to the effects of postmenopausal osteoporosis. This tissue is often referred to as spongy or cancellous bone and is particularly concentrated near the ends of the bone (near the joints) and in the vertebrae of the spine. The trabecular tissue is characterized by small osteoid structures which inter-connect with each other, as well as the more solid and dense cortical tissue which makes up the outer surface and central shaft of the bone. This interconnected network of trabeculae gives lateral support to the outer cortical structure and is critical to the biomechanical strength of the overall structure. In postmenopausal osteoporosis, it is loss of the trabeculae which leads to the failure and fracture of bone. In light of the loss of the trabeculae in postmenopausal women, it is not surprising that the most common fractures are those associated with bones which are highly dependent on trabecular support, e.g., the vertebrae and the neck of the weight bearing bones, such as the femur and the fore-arm. Indeed, hip fracture, collies fractures, and vertebral crush fractures are hallmarks of postmenopausal osteoporosis.

At this time, the generally accepted method for treatment of postmenopausal osteoporosis is estrogen replacement therapy (ERT). Although ERT is generally successful, patient compliance with this therapy is low primarily because estrogen treatment frequently produces undesirable side effects.

Prior to menopause, most women have less incidence of cardiovascular disease than age-matched men. Following menopause, however, the rate of cardiovascular disease in women, such as hyperlipidemia, increases to match the rate seen in men. This rapid increase in the incidence of cardiovascular disease has been linked, in part, to the loss of estrogen and to the loss of estrogen's ability to regulate serum lipids. The nature of estrogen's ability to regulate serum lipids is not well understood, but evidence to date indicates that estrogen can upregulate the low density lipid (LDL) receptors in the liver to remove excess cholesterol. Additionally, estrogen appears to have some effect on the biosynthesis of cholesterol, as well as other beneficial effects on cardiovascular health.

It has been reported in the literature that postmenopausal women undergoing estrogen replacement therapy have a return of serum lipid levels to concentrations similar to those of the premenopausal state. Thus, estrogen would appear to be a reasonable treatment for this condition. However, the side-effects of ERT are not acceptable to many women, thus limiting the use of this therapy. An ideal therapy for this condition would be an agent which would regulate the serum lipid levels like estrogen, but would be devoid of the side-effects and risks associated with estrogen therapy.

The third major pathology associated with, but not limited to, postmenopausal syndrome is estrogen-dependent cancer, primarily breast and uterine cancer. Although such neoplasms are not solely limited to postmenopausal women, they are more prevalent in the older postmenopausal population. Current chemotherapy of these cancers has relied heavily on the use of estrogen agonist/antagonist compounds, such as tamoxifen. Although such mixed agonist/antagonists have beneficial effects in the treatment of these cancers, the estrogenic side-effects are tolerable in only acute life-threatening situations. These agents have stimulatory effects on certain cancer cell populations in the uterus due to their estrogenic (agonist) properties and therefore, are contraproductive in some cases. A better therapy for the treatment of these cancers would be an agent which is an antiestrogenic compound in cancerous tissue, having negligible or no estrogen agonist properties on other reproductive tissues.

In response to the clear need for new pharmaceutical agents which are capable of alleviating the symptoms of, inter alia, postmenopausal syndrome, the present invention provides new compounds, pharmaceutical compositions thereof, and methods of using such compounds for the inhibition of postmenopausal syndrome and other estrogen-related pathological conditions such as those mentioned herein.

SUMMARY OF THE INVENTION

The present invention relates to a compound of formula ##STR2## wherein: m is 0 or 1;

R and R¹ are independently hydroxy, halo, or O--Pg;

X is C═O, CH--OH, CH₂, O, or S;

Pg is independently at each occurrence a hydroxy protecting group; and

R² is substituted C₅ -C₇ cycloalkyl or N-substituted -pyrrolidin-2-yl, -pyrrolidin-3-yl, -piperidin-2-yl, -piperidin-3-yl, -piperidin-4-yl, -hexamethyleneimin-3-yl, or -hexamethyleneimin-4-yl where the N-substituent is C₁ -C₆ alkyl or phenyl(C₁ -C₆ alk-di-yl);

provided that if m is 1, then R² is not substituted C_(5-C) ₇ cycloalkyl, -hexamethyleneimin-3-yl, or -hexamethyleneimin-4-yl, and the N-substituent is not C₁ -C₆ alkyl; or a pharmaceutical salt or solvate thereof.

The present invention further relates to pharmaceutical formulations containing compounds of formula I and the use of such compounds for alleviating the symptoms of postmenopausal syndrome, particularly osteoporosis, cardiovascular-related pathological conditions, and estrogen-dependent cancer.

DETAILED DESCRIPTION OF THE INVENTION

As used herein, the term "C₁ -C₄ alkyl" represents a methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, cyclobutyl, s-butyl, or a t-butyl group. The term "C₁ -C₆ alkyl" includes "C₁ -C₄ alkyl" groups in addition to monovalent, straight, branched, or cyclic saturated hydrocarbons having five or six carbon atoms including, but not limited to, pentyl, isopentyl, hexyl, 2-methylpentyl, cyclopentyl, cyclohexyl, and like groups. The term "C₁ -C₄ alkoxy" refers to a methoxy, ethoxy, propoxy, isopropoxy, butoxy, t-butoxy, and a s-butoxy group. The term "C₅ -C₇ cycloalkyl" refers to a cyclopentyl, cyclohexyl, and a cycloheptyl group.

The term "C₁ -C₆ alk-di-yl" refers to a divalent straight saturated hydrocarbon chain containing from 1 to 6 methylene units.

The term "halo" or "halide" refers to fluoro, chloro, bromo, and iodo.

The term "aryl" refers to a phenyl, substituted phenyl, benzyl, and a substituted benzyl group.

The term "substituted C₅ -C₇ cycloalkyl" refers to a C_(5-C) ₇ cycloalkyl group mono-substituted with a pyrrolidin-1-yl, piperidin-1-yl, or hexamethyleneimin-1-yl group.

An "N-substituted" pyrrolidinyl, piperidinyl, or hexamethyleneiminyl ring is a pyrrolidinyl, piperidinyl, or hexamethyleneiminyl ring respectively substituted at the 1 position of the ring with a C₁ -C₆ alkyl or a phenyl(C₁ -C₆ alk-di-yl) group.

The terms "substituted phenyl" and "substituted benzyl" represent a phenyl and benzyl group respectively substituted with one to three moieties chosen from the group consisting of halo, hydroxy, nitro, C₁ -C₄ alkyl, C₁ -C₄ alkoxy, trichloromethyl, and trifluoromethyl. Examples of a substituted phenyl group include 4-chlorophenyl, 2,6-dichlorophenyl, 2,5-dichlorophenyl, 3,4-dichlorophenyl, 3-chlorophenyl, 3-bromophenyl, 4-bromophenyl, 3,4-dibromophenyl, 3-chloro-4-fluorophenyl, 2-fluorophenyl, 4-hydroxyphenyl, 3-hydroxyphenyl, 2,4-dihydroxyphenyl, 3-nitrophenyl, 4-nitrophenyl, 2,4-dinitrophenyl, 4-methylphenyl, 4-ethylphenyl, 4-methoxyphenyl, 4-propylphenyl, 4-n-butylphenyl, 4-t-butylphenyl, 3-fluoro-2-methylphenyl, 2,3-difluorophenyl, 2,6-difluorophenyl, 2,6-dimethylphenyl, 2-fluoro-5-methylphenyl, 2,4,6-trifluorophenyl, 2-trifluoromethylphenyl, 2-chloro-5-trifluoromethylphenyl, 3,5-bis-(trifluoromethyl)phenyl, 2-methoxyphenyl, 3-methoxyphenyl, 3,5-dimethoxyphenyl, 2-methyl-4-nitrophenyl, 4-methoxy-2-nitrophenyl, and the like. Examples of substituted benzyl groups include all the compounds named when the word "benzyl" is substituted for the word "phenyl" in all the previously mentioned examples of a substituted phenyl group.

The term "hydroxy protecting group" denotes a group understood by one skilled in the organic chemical arts of the type described in Chapter 2 of "protective Groups in Organic Synthesis, 2nd Edition, T. H. Greene, et al., John Wiley & Sons, New York, 1991, hereafter "Greene".

Representative hydroxy protecting groups include, for example, C₁ -C₄ alkyl and substituted C₁ -C₄ alkyl, including methyl, ethyl, or isopropyl, methoxymethyl, methylthiomethyl, t-buylthiomethyl, (phenyldimethylsilyl)methoxymethyl, benzyloxymethyl, p-methoxybenzyloxymethyl, and t-butoxymethyl; ethoxyethyl, 1-(2-chloroethoxy)ethyl, 2,2,2-trichloroethoxymethyl, and 2-(trimethylsilyl)ethyl; phenyl and substituted phenyl groups such as phenyl, p-chlorophenyl, p-methoxyphenyl, and 2,4-dinitrophenyl; benzyl groups; and alkylsilyl groups such as trimethyl- triethyl- and triisopropylsilyl, mixed alkylsilyl groups such as dimethylisopropylsilyl, and diethylisopropylsilyl; acyl protecting groups such as those of the general formula COC₁ -C₆ alkyl or CO(aryl); esters of the general formula CO₂ (C₁ -C₆ alkyl), or CO₂ (aryl), and sulfonyl groups of the general formula SO₂ Ph or SO₂ (C₁ -C₆ alkyl) where Ph is phenyl or substituted phenyl. The species of hydroxy protecting group employed is not critical so long as the derivatized hydroxy group is stable to the condition of subsequent reaction(s) on other positions of the intermediate molecule and can be selectively removed at the appropriate point without disrupting the remainder of the molecule including any other hydroxy protecting group(s). It is within the knowledge of one skilled in the art to select appropriate hydroxy protecting group(s) for a given set of reaction conditions given the guidance provided by Greene cited above.

Although the free-base form of formula I compounds can be used in the methods of the present invention, it is preferred to prepare and use a pharmaceutical salt form. Typical pharmaceutical salts include those salts prepared by reaction of the compounds of the present invention with a mineral or organic acid. Such salts are known as acid addition salts. Thus, the term "pharmaceutical salt" refers to acid addition salts of a compound of formula I which are substantially non-toxic at the doses administered and are commonly known in the pharmaceutical literature. See e.g. Berge, S. M, Bighley, L. D., and Monkhouse, D. C., J. Pharm. Sci., 66, 1, 1977.

Examples of such pharmaceutically acceptable salts are the iodide, acetate, phenylacetate, trifluoroacetate, acrylate, ascorbate, benzoate, chlorobenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, methylbenzoate, o-acetoxybenzoate, naphthalene-2-benzoate, bromide, isobutyrate, phenylbutyrate, g-hydroxybutyrate, b-hydroxybutyrate, butyne-1,4-dioate, hexyne-1,4-dioate, hexyne-1,6-dioate, caproate, caprylate, chloride, cinnamate, citrate, decanoate, formate, fumarate, glycollate, heptanoate, hippurate, lactate, malate, maleate, hydroxymaleate, malonate, mandelate, mesylate, nicotinate, isonicotinate, nitrate, oxalate, phthalate, terephthalate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, propiolate, propionate, phenylpropionate, salicylate, sebacate, succinate, suberate, sulfate, bisulfate, pyrosulfate, sulfite, bisulfite, sulfonate, benzenesulfonate, p-bromophenylsulfonate, chlorobenzenesulfonate, propanesulfonate, ethanesulfonate, 2-hydroxyethanesulfonate, methanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, p-toluenesulfonate, xylenesulfonate, tartarate, and the like of a compound of formula I.

By "pharmaceutical formulation" it is meant that in a formulation containing the compound of formula I, the carrier, diluent, excipients, and salt must be compatible with the other ingredients of the formulation, and not deleterious to the recipient thereof.

The term "solvate" represents an aggregate that comprises one or more molecules of the solute, such as a formula I compound, with one or more molecules of solvent.

As used herein, the term "effective amount" means an amount of compound of the present invention which is capable of inhibiting the symptoms of the various pathological conditions herein described.

The terms "inhibit" or inhibiting bear their usual meaning which includes prohibiting, treating, alleviating, ameliorating, halting, restraining, slowing or reversing the progression, or reducing the severity of a pathological symptom related to or resultant from post menopausal syndrome. As such, these methods include both medical therapeutic (acute) and/or prophylactic (prevention) administration as appropriate.

The R² moiety contains a chiral center, thus, when X is not CH--OH, the compounds of formula I are enantiomeric. When X is CH--OH, the compounds of formula I are diastereomeric. Single enantiomers, single diastereomers, and mixtures id thereof, are included within the scope of the present invention. While all diastereomers, enantiomers, and mixtures thereof, are useful, single enantiomers or single diastereomers are preferred.

While all of the compounds of the present invention are useful, certain of the compounds are particularly interesting and are preferred. The following listing sets out several groups of preferred compounds. It will be understood that each of the listings may be combined with other listings to create additional groups of preferred compounds.

aa) m is 0;

ab) m is 1;

ac) Pg is C₁ -C₄ alkyl;

ad) Pg is benzyl;

ae) Pg is isopropyl;

af) Pg is methyl;

ag) R and R¹ are independently hydroxy or O--Pg;

ah) R² is N-substituted-piperidin-2-yl;

ai) R² is N-substituted-piperidin-4-yl;

aj) R² is N-substituted hexamethyleneimin-3-yl;

ak) R² is cyclohexyl substituted with piperidin-1-yl;

al) R² is cyclohexyl substituted at the 2 position;

am) N-substituted is N-methyl;

an) N-substituted is N-ethyl;

ao) N-substituted is N-n-hexyl;

ap) N-substituted is N-benzyl;

aq) X is C═O;

ar) X is O;

as) the compound of formula I is a salt;

at) the compound of formula I is the hydrochloride salt.

Specific preparations of compounds of the present invention are described herein, in Examples 1-30. Modification to the methods described below may be necessary to accommodate reactive functionalities of particular substituents. Such modification would be both apparent to, and readily ascertained by, those skilled in the art. The following schemes generally illustrate the preparation of compounds of formula I.

The compounds of formula I may be prepared from compounds of formula II as illustrated in Scheme 1 below where R' and R^(1') are independently at each occurrence halo, or O--Pg, R², is substituted C₅ -C₇ cycloalkyl or N-substituted -pyrrolidin-2-yl, -pyrrolidin-3-yl, -piperidin-2-yl, -piperidin-3-yl, or -piperidin-4-yl, X' is C═O, O, or S, and m and R² are as described supra. ##STR3##

Compounds of formula I(a) may be prepared by the Mitsunobo reaction of a compound of formula II and III. This transformation is accomplished by dissolving or suspending a compound of formula II in a suitable solvent and adding a compound of formula III, triphenyl phosphine, and diethylazodicarboxylate. The resulting mixture is allowed to stir for from 2 to 24 hours, but the reaction is typically complete in from 16 to 20 hours. The reaction is preferably allowed to stir for about 18 hours. Suitable solvents include anhydrous solvents, such as methylene chloride, acetonitrile, chloroform, ethyl acetate, mixtures thereof, and the like. Typically, anhydrous tetrahydrofuran is a convenient and preferred solvent. The compound of formula III, triphenyl phosphine, and diethylazodicarboxylate are typically employed in a molar excess relative to the compound of formula II. The compounds of formula III, triphenyl phosphine, and diethylazodicarboxylate are usually employed in about a 1.8 to about a 2.2 molar excess while a 2.0 molar excess is typically preferred. For further instruction on conditions and reagents useful in the Mitsunobo reaction see Mitsunobo's review article in Synthesis, 1, (1981).

When the Mitsunobo reaction is performed as described above with a compound of formula III where R^(2') is N-substituted -piperidin-2-yl, -piperidin-3-yl, or -piperidin-4-yl, a mixture of products results. A component of this mixture is the expected corresponding compound of formula I(a) where R² is N-substituted -piperidin-2-yl, -piperidin-3-yl, or -piperidin-4-yl. In addition, as a result of a ring expansion that occurs under these Mitsunobo conditions, another component of the product mixture is a compound of formula I(a) where R² is a hexamethyleneimin-3-yl or a hexamethyleneimin-4-yl ring. The correspondence between starting materials and products is illustrated in Table I below where R³ is C₁ -C₆ alkyl or phenyl(C₁ -C₆ alk-di-yl).

                                      TABLE 1                                      __________________________________________________________________________     When the compound of formula III is                                                             R.sup.2 in the compound of formula I(a) will be               __________________________________________________________________________                        #STR4##                                                                        #STR5##                                                        -                                                                                              #STR6##                                                                        #STR7##                                                        -                                                                                              #STR8##                                                                        #STR9##                                                     __________________________________________________________________________

The ratio of 6 to 7 membered cyclic product is typically about 1:1. The two products may be separated and purified by conventional chromatographic techniques. Thus, compounds of formula I where R² is a hexamethyleneiminyl ring, may be accessed by the Mitsunobo reaction of a compound of formula II with a N-substituted piperidin-2-yl methanol, piperidin-3-yl methanol, or piperidine-4-yl methanol of formula III. For further illustration of this ring expansion chemistry and the resulting products, see Examples 5-8.

The compounds of formula I where X is CH--OH or CH₂ may be prepared from compounds of formula I where X is C═O essentially as described in U.S. Pat. No. 5,484,798, the teachings of which are hereby incorporated by reference.

When any of R, R', R¹, or R^(1') are hydroxy protecting groups in compounds of formula I or I(a), they may be removed by well known methods in the art to give the compounds of formula I where R and or R¹ are hydroxy. Numerous reactions for the installation and removal of hydroxy protecting groups are described in a number of standard works including, for example, Protective Groups in Organic Chemistry, Plenum Press (London and New York, 1973); The Peptides, Vol. I, Schrooder and Lubke, Academic Press (London and New York, 1965), and Greene. Preferred protecting groups are benzyl and C₁ -C₄ alkyl groups and especially preferred are methyl and isopropyl groups.

The pharmaceutical acid addition salts are typically formed by reacting a compound of formula I in its free base form with an equimolar or excess amount of acid. The reactants are generally combined in a polar organic solvent such as methanol or ethyl acetate. The salt normally precipitates out of solution within about one hour to 10 days and can be isolated by filtration, or the solvent can be stripped off by conventional means.

The pharmaceutical salts generally have enhanced solubility characteristics compared to the compound from which they are derived, and thus are often more amenable to use in pharmaceutical formulations.

The compounds of formula II may be prepared by a number of well known routes. For example, compounds of formula II where X is C═O and compounds of formula II where X is O, or S can be prepared from m-methoxythiophenol and an appropriately substituted a-bromoacetophenone as taught respectively in U.S. Pat. Nos. 4,133,814 and 5,510,357, the teachings of which are hereby incorporated by reference.

Compounds of formula III are either commercially available or may be prepared by methods well known in the art.

The optimal time for performing the reaction of Scheme 1 can be determined by monitoring the progress of the reaction via conventional chromatographic techniques. Furthermore, it is preferred to conduct the reactions of the invention under an inert atmosphere, such as, for example, argon, or, particularly, nitrogen. Choice of solvent is generally not critical so long as the solvent employed is inert to the ongoing reaction and sufficiently solubilizes the reactants to effect the desired reaction. Intermediates and final products may be purified, if desired by common techniques such as recrystallization or chromatography over solid supports such as silica gel or alumina.

The synthetic steps of the routes described herein may be combined in other ways to prepare the formula I compounds. The discussion of the synthesis is not intended to be limiting to the scope of the present invention, and should not be so construed. Application of the above chemistry enables the synthesis of the compounds of formula I, which include, but are not limited to:

6-methoxy-2-(4-methoxyphenyl)-3-(4-[1-hexylhexamethyleneimin-3-oxy]benzoyl)benzo[b]thiophene;

6-hydroxy-2-(4-isopropoxyphenyl)-3-(4-[1-benzylpiperidin-3-oxy]benzoyl)benzo[b]thiophene;

6-hydroxy-2-(4-hydroxyphenyl)-3-(4-[1-propylpyrrolidin-3-oxy]benzoyl)benzo[b]thiophene;

6-benzyloxy-2-(4-isopropoxyphenyl)-3-(4-[1-hexylhexamethyleneimin-2-oxy]benzoyl)benzo[b]thiophene;

6-chloro-2-(4-chlorophenyl)-3-(4-[1-methylpiperidin-2-oxy]benzoyl)benzo[b]thiophene;

6-fluoro-2-(4-fluorophenyl)-3-(4-[1-butylpyrrolidin-2-oxy)benzoyl)benzo[b]thiophene;

6-bromo-2-(4-bromophenyl)-3-(4-[1-pentylhexamethyleneimin-4-oxybenzoyl)benzo[b]thiophene;

6-methoxy-2-(4-methoxyphenyl)-3-(4-[1-Phenethylpiperidin-4-oxy]benzoyl)benzo[b]thiophene;

6-hydroxy-2-(4-isopropoxyphenyl)-3-(4-[3-pyrrolidin-1-ylcycloheptoxy]benzoyl)benzo[b]thiophene;

6-hydroxy-2-(4-hydroxyphenyl)-3-(4-[2-piperidin-1-ylcyclopentoxy]benzoyl)benzo[b]thiophene;

6-benzyloxy-2-(4-isopropoxyphenyl)-3-(4-[3-hexamethyleneimin-1-ylcyclohexoxy]benzoyl)benzo[b]thiophene;

6-hydroxy-2-(4-isopropoxyphenyl)-3-(4-[(1-benzylpiperidin-3-yl)methoxy]benzoyl)benzo[b]thiophene;

6-hydroxy-2-(4-hydroxyphenyl)-3-(4-[(1-propylpyrrolidin-3-yl)methoxy]benzoyl)benzo[b]thiophene;

6-chloro-2-(4-chlorophenyl)-3-(4-[(1-methylpiperidin-2-yl)methoxy]benzoyl)benzo[b]thiophene;

6-fluoro-2-(4-fluorophenyl)-3-(4-[(1-butylpyrrolidin-2-yl)methoxy]benzoyl)benzo[b]thiophene;

6-methoxy-2-(4-methoxyphenyl)-3-(4-[(1-phenethylpiperidin-4-yl)methoxy]benzoyl)benzo[b]thiophene;

6-hydroxy-2-(4-isopropoxyphenyl)-3-(4-[(3-pyrrolidin-1-ylcycloheptyl)methoxy]benzoyl)benzo[b]thiophene;

6-hydroxy-2-(4-hydroxyphenyl)-3-(4-[(2-piperidin-1-ylcyclopentyl)methoxy]benzoyl)benzo[b]thiophene; and

6-benzyloxy-2-(4-isopropoxyphenyl)-3-(4-[(3-hexamethyleneiminylcyclohexyl)methoxy]benzoyl)benzo[b]thiophene.

The following Preparations and Examples further illustrate the synthesis of the compounds of the present invention. The examples are not intended to be limiting to the scope of the invention in any respect, and should not be so construed. All experiments were run under positive pressure of dry nitrogen. The terms and abbreviations used in the instant preparations and examples have their normal meanings unless otherwise designated. For example "OC", "N", "mmol", "g", "mL", "M", "HPLC", "EA", "IR" and "¹ H-NMR", refer to degrees Celsius, normal or normality, millimole or millimoles, gram or grams, milliliter or milliliters, molar or molarity, high performance liquid chromatography, elemental analysis, infrared, and proton nuclear magnetic resonance respectively. In addition, the absorption maxima listed for the IR spectra are only those of interest and not all of the maxima observed.

PREPARATIONS Preparation 1 4-Hydroxy-1-Hexylpiperidine

4-Hydroxypiperidine (2.02 g, 20 mmol) was dissolved in 100 mL of methyl ethylketone. To this stirred mixture was added 2-iodohexane (1.5 mL, 10 mmol) and the resulting solution was heated at reflux for 1.5 hours. The mixture was allowed to cool and was then washed with saturated aqueous sodium bicarbonate (3×75 mL). The organic layer was dried over magnesium sulfate, filtered, and evaporated, and the residue was dried overnight under house vacuum to yield 1.184 g (64%) of the title compound which was used without further purification. ¹ H NMR consistent with desired product. MS(FD) 186(M+).

Preparation 2 1-Hexyl-2-Piperidine Methanol

2-Piperidine methanol (2.30 g, 20 mmol) and 1-iodohexane (1.5 mL, 10 mmol) were converted to product by the procedure of Preparation 1 to yield 2.048 g (51%) of the title compound. ³ H NMR consistent with desired product.

Preparation 3 1-Hexyl-3-Piperidine Methanol

3-Piperidine methanol (2.30 g, 20 mmol) and 1-iodohexane (1.5 mL, 10 mmol) were converted to product by the procedure of Preparation 1 to yield 1.866 g (47%) of the title compound which was used without further purification. ¹ H NMR consistent with desired product.

Preparation 4 1-(2-Hydroxycyclohexyl )pyrrolidine

Cyclohexene oxide (982 mg, 10 mmol), pyrolidine (2.134 g, 30 mmol), and 1.8 mL of water were heated at 90° C. for 18 hours. The reaction was then cooled and diluted with 75 mL of diethyl ether. The mixture was then washed with aqueous sodium bicarbonate (15 mL), brine (2×15 mL), dried over magnesium sulfate, filtered, and evaporated to give 1.499 g (89%) of the title compound.

Preparation 5 1-(2-Hydroxycyclohexyl)piperidine

Piperidine hydrochloride (3.648 g, 30 mmol) was stirred in a mixture of 4.0 mL of water and 4.146 g of ground potassium carbonate. This mixture was heated to 90° C. and cyclohexene oxide (1.01 mL, 10 mmol) was added. The resulting mixture was heated at 100° C. for 18 hours. The reaction was cooled then filtered to remove carbonate salts. The filter cake was washed with 25 mL of diethyl ether and 50 mL of ethyl acetate. The filtrate and washings were combined and washed with brine (2×20 mL), dried over magnesium sulfate, filtered, and evaporated to give 1.875 g (>100%) of the title compound which was used without further purification. ¹ H NMR consistent with desired product.

EXAMPLES Example 1 6-Methoxy-2-(4-Methoxyphenyl)-3-(4-[1-Methylpiperidin-4-oxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-hydroxybenzoyl)benzo[b]thiophene (1.17 g, 3.00 mmol) and 4-hydroxy-1-methylpiperidine (691 mg, 6.00 mmol) were dissolved in 50 mL of anhydrous tetrahydrofuran. This solution was stirred and triphenylphosphine (1.57 g, 6.00 mmol) and diethylazodicarboxylate (DEAD) (6.00 mmol) were added. The resulting solution was allowed to stir at room temperature for 18 hours. The solvent was removed under reduced pressure and the resulting mixture is flash chromatographed (eluent: 7:3 ethyl acetate:methanol) to give 834 mg of the title compound. Yield: 57%. MS(FD) 488(M+). EA calculated for C₂₉ H₂₉ NO₄ S: C, 71.43; H, 5.99; N, 2.87. Found: C, 71.14; H, 5.93; N, 2.61.

Example 2 6-Methoxy-2-(4-Methoxyphenyl)-3-(4-[1-Ethylpiperidin-4-oxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-hydroxybenzoyl)benzo[b]thiophene (1.17 g, 3.00 mmol), 4-hydroxy-1-ethylpiperidine (775 mg, 6.00 mmol), triphenylphosphine (1.57 g, 6.00 mmol), and DEAD (6.00 mmol) were converted to product by the procedure of Example 1 to give 827 mg of the title compound. Yield: 55%. MS(FD) 501(M+). EA calculated for C₃₀ H₃₁ NO₄ S: C, 71.83; H, 6.23; N, 2.79. Found: C, 71.61; H, 5.94; N, 2.69.

Example 3 6-Methoxy-2-(4-Methoxyphenyl)-3-(4-[1-Benzylpiperidin-4-oxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-hydroxybenzoyl)benzo[b]thiophene (1.17 g, 3.00 mmol), 4-hydroxy-1-benzylpiperidine (1.15 g, 6.00 mmol), triphenylphosphine (1.57 g, 6.00 mmol), and DEAD (6.00 mmol) were converted to product by the procedure of Example 1 to give 1.33 g of the title compound. Yield: 79%. MS(FD) 563(M+). IR (CHCl₃) ν max 3030, 3011, 2955, 2846, 1645, 1597, 1476, 1254, 1166.

Example 4 6-Methoxy-2-(4-Methoxyphenyl)-3-(4-[1-Hexylpiperidin-4-oxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-hydroxybenzoyl)benzo[b]thiophene (895 mg, 2.30 mmol), 4-hydroxy-1-hexylpiperidine (850 mg, 4.59 mmol), triphenylphosphine (1.20 g, 4.59 mmol), and DEAD (4.59 mmol) were converted to product by the procedure of Example 1 to give 653 mg of the title compound. Yield: 51%. MS(FD) 557(M+). EA calculated for C₃₄ H₃₉ NO₄ S: C, 73.21; H, 7.05; N, 2.51. Found: C, 73.46; H, 7.26; N, 2.57.

Examples 5 and 6 6-Methoxy-2-(4-Methoxyphenyl)-3-(4-[(1-Methylpiperidin-2-yl)methoxy]benzoyl)benzo[b]thiophene and 6-Methoxy-2-(4-Methoxyphenyl)-3-(4-[1-Methylhexamethyleneimin-3-oxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-hydroxybenzoyl)benzo[b]thiophene (1.17 g, 3.00 mmol), 1-methyl-2-piperidine methanol (775 mg, 6.00 mmol), triphenylphosphine (1.57 g, 6.00 mmol), and DEAD (6.00 mmol) were converted to product by the procedure of Example 1 to give 496 mg (33%) of the piperidinyl title compound; MS(FD) 501(M+); EA calculated for C₃₀ H₃₁ NO₄ S: C, 71.83; H, 6.23; N, 2.79. Found: C, 72.07; H, 6.40; N, 2.69; and 1.08 g (38%) of the ring expanded title compound. MS(FD) 571(M+). IR (CHCl₃) ν max 3031, 3010, 2941, 2850, 1646, 1597, 1476, 1253, 1179, 1166.

Examples 7 and 8 6-Methoxy-2-(4-Methoxyphenyl)-3-(4-[(1-Hexylpiperidin-2-yl)methoxy]benzoyl)benzo[b]thiophene and 6-Methoxy-2-(4-methoxyphenyl)-3-(4-[1-Hexylhexamethyleneimin-3-oxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-hydroxybenzoyl)benzo[b]thiophene (913 mg, 2.34 mmol), 1-hexyl-2-piperidine methanol (1.66 g, 5.80 mmol), triphenylphosphine (1.23 g, 4.7 mmol), and DEAD (4.7 mmol) were converted to product by the procedure of Example 1 to give 616 mg (46%) of the piperidinyl title compound; MS(FD) 572(M+); IR (CHCl₃) ν max 3015, 2971, 2937, 1646, 1600, 1476, 1254, 1167; and 468 mg (35%) of the ring expanded title compound. MS(FD) 571(M+). IR (CHCl₃) ν max 3009, 2936, 2861, 1646, 1597, 1476, 1253, 1166.

Example 9 6-Methoxy-2-(4-Methoxyphenyl)-3-(4-[(1-Hexylpiperidin-3-yl)methoxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-hydroxybenzoyl)benzo[b]thiophene (1.17 g, 3.00 mmol), 1-hexyl-3-piperidine methanol (1.69 g, 6.00 mmol), triphenylphosphine (1.57 g, 6.00 mmol), and DEAD (6.0 mmol) were converted to product by the procedure of Example 1 to give 1.41 g of the title compound. Yield: 82%. MS(FD) 572(M+). EA calculated for C₃₅ H₄₁ NO₄ S: C, 73.52; H, 7.23; N, 2.45. Found: C, 73.25; H, 7.08; N, 2.71.

Example 10 6-Methoxy-2-(4-Methoxyphenyl)-3-(4-[1-Benzylpyrrolidin-3-oxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-hydroxybenzoyl)benzo[b]thiophene (1.17 g, 3.00 mmol), 1-benzyl-3-hydroxypyrrolidine (1.06 g, 6.00 mmol), triphenyiphosphine (1.57 g, 6.00 mmol), and DEAD (6.0 mmol) were converted to product by the procedure of Example 1 to g ive 1.57 g of the title compound. Yield: 95.3%. MS(FD) 549(M+). IR (CHCl₃) ν max 3033, 3010, 2971, 2842, 1646, 1599, 1476, 1254, 1167.

Example 11 6-Methoxy-2-(4-Methoxyphenyl)-3-(4-[(1-Methylpiperidin-3-yl) methoxy]benzoyl) benzo [b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-hydroxybenzoyl)benzo[b]thiophene (1.17 g, 3.00 mmol) and 1-methyl-piperidine methanol (0.775 g, 6.00 mmol) were dissolved in anhydrous tetrahydrofuran (50 mL). To this stirred solution was added triphenylphosphine (1.57 g, 6.00 mmol) followed by diethylazodicarboxylate (0.945 mL, 6.00 mmol) via syringe, and the reaction mixture was stirred at room temperature for 18 hours. The solvent was removed under reduced pressure, and the resulting mixture was flash chromatographed (eluent 7:3 ethyl acetate:methanol) to give 1.176 g (78%) of title compound. MS(FD) 501(M+). IR (CHCl₃) ν max 3030, 3011, 2941, 2846, 2795, 1646, 1599, 1476, 1254, 1166.

Example 12 6-Methoxy-2-(4-Methoxyphenyl)-3-(4-[1-Methylpiperidin-3-oxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-hydroxybenzoyl)benzo[b]thiophene (1.17 g, 3.00 mmol), 1-methyl-3-hydroxypiperidine (691 mg, 6.00 mmol), triphenylphosphine (1.57 g, 6.00 mmol), and DEAD (6.0 mmol) were converted to product by the procedure of Example 11 to give 307 mg of the title compound. Yield: 21%. MS(FD) 487(M+). IR (CHCl₃) ν max 3010, 2947, 2840, 2795, 1646, 1599, 1476, 1254, 1167.

Example 13 6-Methoxy-2-(4-Methoxyphenyl)-3-(4-[1-Ethylpiperidin-3-oxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-hydroxybenzoyl)benzo[b]thiophene (1.17 g, 3.00 mmol), 1-ethyl-3-hydroxypiperidine (775 mg, 6.00 mmol), triphenylphosphine (1.57 g, 6.00 mmol), and DEAD (6.0 nmol) were converted to product by the procedure of Example 11 to give 256 mg of the title compound. Yield: 17%. MS(FD) 501(M+). EA calculated for C₃₀ H₃₁ NO₄ S: C, 71.83; H, 6.23; N, 2.79. Found: C, 72.10; H, 6.47; N, 3.07.

Example 14 6-Methoxy-2-(4-Methoxyphenyl)-3-(4-[2-Piperidin-1-ylcyclohexoxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-hydroxybenzoyl)benzo[b]thiophene (1.17 g, 3.00 mmol), trans-2-piperidinylcyclohexanol (1.10 g, 6.00 mmol), triphenylphosphine (1.57 g, 6.00 mmol), and DEAD (6.0 mmol) were converted to product by the procedure of Example 11 to give 1.08 g of the title compound. Yield: 65%. MS(FD) 555(M+). IR (CHCl₃) ν max 3014, 2938, 2860, 1644, 1595, 1476, 1254, 1165.

Example 15 6-Methoxy-2-(4-Methoxyphenyl)-3-(4-[2-Pyrrolidin-1-ylcyclohexoxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-hydroxybenzoyl)benzo[b]thiophene (1.27 g, 3.25 mmol), trans-2-pyrrolidinylcyclohexanol (1.10 g, 6.50 mmol), triphenylphosphine (1.70 g, 6.50 mmol), and DEAD (6.50 mmol) were converted to product by the procedure of Example 11 to give 615 mg of the title compound. Yield: 35%. MS(FD) 541(M+). IR (CHCl₃) ν max 3009, 2940, 1645, 1597, 1476, 1254, 1166.

Example 16 6-Hydroxy-2-(4-Hydroxyphenyl)-3-(4-[1-Methylpiperidin-4-oxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-[1-methylpiperidin-4-oxy]benzoyl)benzo[b]thiophene (749 mg, 1.54 mmol) was dissolved in 30 mL of methylene chloride. This solution was stirred and ethane thiol (7.68 mmol) and aluminum chloride (1.23 g, 9.22 mmol) were added. The reaction mixture was stirred vigorously for 30 minutes and then quenched with brine and saturated sodium bicarbonate. Any resulting residue was dissolved by the addition of methanol and ethyl acetate. The pH was adjusted to make it slightly basic and the mixture was diluted with 150 mL of ethyl acetate. After separation of the aqueous layer, the organic layer was washed with 75 mL of aqeous potassium sodium tartrate. The organic layer was dried over magnesium sulfate, filtered, and evaporated under reduced pressure. The residue was purified by rotary chromatography (4 mm. Plate, eluting solvent 4:1 ethyl acetate:methanol) to give 452 mg (64%) of the title compound as a yellow solid. MS(FD) 459(M+). IR (CHCl₃) ν max 3221, 2960, 1598, 1467, 1241, 1166.

Example 17 6-Hydroxy-2-(4-Hydroxyphenyl)-3-(4-[1-Ethylpiperidin-4-oxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-[1-ethylpiperidin-4-oxy]benzoyl)benzo[b]thiophene (502 mg, 1.00 mmol), ethane thiol (5.00 mmol), and aluminum chloride (800 mg, 6.00 mmol) were converted to 341 mg (72%) of the title compound by the procedure of Example 16. MS(FD) 571(M+). IR (CHCl₃) ν max 3300, 2971, 1598, 1249, 1165.

Example 18 6-Hydroxy-2-(4-Hydroxyphenyl)-3-(4-[1-Benzylpiperidin-4-oxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-[1-benzylpiperidin-4-oxy]benzoyl)benzo[b]thiophene (846 mg, 1.50 mmol), ethane thiol (7.50 mmol), and aluminum chloride (1.2 g, 9.00 mmol) were converted to 450 mg (56%) of the title compound by the procedure of Example 16. MS(FD) 536(M+). IR (CHCl₃) ν max 3311, 3023, 3011, 2953, 1721, 1596, 1469, 1257, 1166.

Example 19 6-Hydroxy-2-(4-Hydroxyphenyl)-3-(4-[1-Hexylpiperidin-4-oxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-[1-hexylpiperidin-4-oxy]benzoyl)benzo[b]thiophene (375 mg, 0.941 mmol), ethane thiol (3.36 mmol), and aluminum chloride (538 mg, 4.04 mmol) were converted to 209 mg (42%) of the title compound by the procedure of Example 16. MS(FD) 529(M+). EA calculated for C₃₂ H₃₅ NO₄ S: C, 72.56; H, 6.66; N, 2.64. Found: C, 72.33; H, 6.89; N, 2.52.

Example 20 6-Hydroxy-2-(4-Hydroxyphenyl)-3-(4-[(1-Methylpiperidin-2-yl)methoxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-[(1-methylpiperidin-2-yl)methoxy]benzoyl)benzo[b]thiophene (339 mg, 0.676 mmol), ethane thiol (8.75 mmol), and aluminum chloride (541 mg, 4.06 mmol) were converted to 195 mg (61%) of the title compound by the procedure of Example 16. MS(FD) 473(M+). IR (CHCl₃) ν max 3414, 3179, 1605, 1468, 1261, 1170.

Example 21 6-Hydroxy-2-(4-Hydroxyphenyl)-3-(4-[(1-Hexylpiperidin-2-yl)methoxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-[(1-hexylpiperidin-2-yl)methoxy]benzoyl)benzo[b]thiophene (464 mg, 0.812 mmol), ethane thiol (4.06 mmol), and aluminum chloride (649 mg, 4.87 mmol) were converted to 194 mg (44%) of the title compound by the procedure of Example 16. MS(FD) 544(M+). IR (CHCl₃) ν max 3380, 2930, 2858, 1598, 1467, 1256, 1166.

Example 22 6-Hydroxy-2-(4-Hydroxyphenyl)-3-(4-[(1-Hexylpiperidin-3-yl)methoxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-[(1-hexylpiperidin-3-yl)methoxy]benzoyl)benzo[b]thiophene (1.24 g, 2.17 mmol), ethane thiol (10.8 mmol), and aluminum chloride (1.73 g, 13.0 mmol) were converted to 224 mg (19%) of the title compound by the procedure of Example 16. MS(FD) 544(M+). EA calculated for C₃₃ H₃₇ NO₄ S: C, 72.90; H, 6.86; N, 2.58. Found: C, 73.14; H, 7.04; N, 2.52.

Example 23 6-Hydroxy-2-(4-Hydroxyphenyl)-3-(4-[1-Benzylpyrrolidin-3-oxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-[1-benzylpyrrolidin-3-oxy]benzoyl)benzo[b]thiophene (1.57 g, 2.86 mmol), ethane thiol (14.3 mmol), and aluminum chloride (2.29 g, 17.2 mmol) were converted to 566 mg (38%) of the title compound by the procedure of Example 16. MS(FD) 521(M+). IR (CHCl₃) ν max 3300, 3027, 3012, 2981, 1732, 1599, 1469, 1257, 1167.

Example 24 6-Hydroxy-2-(4-Hydroxyphenyl)-3-(4-[(1-Methylpiperidin-3-yl)methoxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-[(1-methylpiperidin-3-yl)methoxy]benzoyl)benzo[b]thiophene (878 mg, 1.75 mmol), ethane thiol (8.75 mmol), and aluminum chloride (1.40 g, 10.S mmol) were converted to 447 mg (54%) of the title compound by the procedure of Example 16. MS(FD) 473(M+). IR (CHCl₃) ν max 3205, 2954, 1599, 1467, 1266, 1171.

Example 25 6-Hydroxy-2-(4-Hydroxyphenyl)-3-(4-[1-Methylpiperidin-3-oxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-[1-methylpiperidin-3-oxy]benzoyl)benzo[b]thiophene (310 mg, 0.636 mmol), ethane thiol (3.18 mmol), and aluminum chloride (508 mg, 3.81 mmol) were converted to 211 mg (72%) of the title compound by the procedure of Example 16. MS(FD) 460(M+). IR (CHCl₃) ν max 3400, 3149, 2947, 1598, 1467, 1254, 1164.

Example 26 6-Hydroxy-2-(4-Hydroxyphenyl)-3-(4-[2-Ethylpiperidin-3-oxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-[2-ethylpiperidin-3-oxy]benzoyl)benzo[b]thiophene (226 mg, 0.451 mmol), ethane thiol (2.25 mmol), and aluminum chloride (360 mg, 2.70 mmol) were converted to 192 mg (90%) of the title compound by the procedure of Example 16. MS(FD) 474(M+). IR (CHCl₃) ν max 3380, 2970, 1597, 1467, 1255, 1165.

Example 27 6-Hydroxy-2-(4-Hydroxyphenyl)-3-(4-[2-piperidin-3-ylcyclohexoxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-[2-piperidin-1-ylcyclohexoxy]benzoyl)benzo[b]thiophene (562 mg, 1.04 mmol), ethane thiol (5.19 mmol), and aluminum chloride (829 mg, 6.22 mmol) were converted to 481 mg (90%) of the title compound by the procedure of Example 16. MS(FD) 514(M+). IR (CHCl₃) ν max 3386, 2935, 2861, 1595, 1256, 1165.

Example 28 6-Hydroxy-2-(4-Hydroxyphenyl)-3-(4-[2-Pyrrolidin-1-ylcyclohexoxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-[2-pyrrolidin-1-ylcyclohexoxy]benzoyl)benzo[b]thiophene (610 mg, 1.10 mmol), ethane thiol (5.50 mmol), and aluminum chloride (880 mg, 6.60 mmol) were converted to 523 mg (90%) of the title compound by the procedure of Example 16. MS(FD) 528(M+). IR (CHCl₃) ν max 3377, 2934, 2857, 1595, 1467, 1258, 1164.

Example 29 6-Hydroxy-2-(4-Hydroxyphenyl)-3-(4-[1-Methylhexamethyleneimin-3-oxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-[1-methylhexamethyleneimin-3-oxy]benzoyl) benzo [b]thiophene (472 mg, 0.941 mmol), ethane thiol (4.70 inmol), and aluminum chloride (753 mg, 5.65 mmol) were converted to 361 mg (81%) of the title compound by the procedure of Example 16. MS(FD) 474(M+). IR (CHCl₃) ν max 3300, 3008, 2977, 2941, 1598, 1469, 1256, 1168.

Example 30 6-Hydroxy-2-(4-Hydroxyphenyl)-3-(4-[1-Hexylhexamethyleneimin-3-oxy]benzoyl)benzo[b]thiophene

6-Methoxy-2-(4-methoxyphenyl)-3-(4-[1-hexylhexamethyleneimin-3-oxy]benzoyl)benzo[b]thiophene (396 mg, 0.693 mmol), ethane thiol (3.46 mmol), and aluminum chloride (554 mg, 4.16 mmol) were converted to 290 mg (77%) of the title compound by the procedure of Example 16. MS(FD) 544(M+). IR (CHCl₃) ν max 3300, 3010, 2934, 2861, 1597, 1469, 1259, 1166.

Representative compounds of the present invention have been biologically tested to demonstrate their efficacy for inhibiting the effects of post menopausal syndrome.

Estrogenicity: Four Day Ovariectomized Rat Model

Seventy-five day old female Sprague Dawley rats (weight range of 225 g-275 g) were obtained from Charles River Laboratories (Portage, Mich.). The animals were either bilaterally ovariectomized (OVX) or exposed to a Sham surgical procedure at Charles River Laboratories, and then shipped after one week. Upon arrival, these rats were housed in metal hanging cases in groups of three or four animals per cage, and had ad libitum access to food (Teklad diet, TD 89222, 0.5% calcium, 0.4% phosphorous; Madison, Wis.) and water. Room temperature was maintained at 22.2° C. +1.7° C. with a minimum relative humidity of 40%. The photoperiod in the room was twelve hours light and twelve hours dark. Comparative data were obtained between untreated ovariectomized rats, ovariectomized rats treated with 17a-ethynylestradiol (EE₂), and ovariectomized rats treated with representative compounds of the present invention.

After a one-week acclimation period (two weeks post-OVX), daily dosing with test compound and 17a-ethynylestradiol was initiated. The test compounds or 17a-ethynylestradiol (Sigma Chemical Co., St. Louis, Mo.) were given orally, unless otherwise stated, as a suspension in 1% carboxymethylcellulose or dissolved in 20% cyclodextrin. Animals were dosed daily for four days. Following the dosing regimen, animals were weighed and anesthetized with a ketamine:xylazine (2:1, v:v) mixture and a blood sample was collected by cardiac puncture. The animals were then sacrificed by asphyxiation with CO₂, the uterus was removed through a midline incision, and a wet uterine weight was determined.

Serum Cholesterol Analysis

The blood samples, collected as described above, were allowed to clot at room temperature for two hours, and serum was obtained following centrifugation for ten minutes at 3000 rpm. Serum cholesterol was determined using a high-performance cholesterol assay (Boehringer Mannheim Diagnostics, Indianapolis, IN). Briefly, the cholesterol was oxidized to produce cholest-4-en-3-one and hydrogen peroxide. The hydrogen peroxide was then reacted with phenol and 4-aminophenazone in the presence of peroxidase to produce a p-quinoneimine dye, which was read spectrophotometrically at 500 nm. Cholesterol concentration 2 was then calculated against a standard curve.

Uterine Eosinophil Peroxidase (EPO) Assay

Uteri were kept at 4° C. until time of enzymatic analysis. The uteri were then homogenized in 50 volumes of 50 nM Tris buffer (pH=8.0) containing 0.005% Triton X-100. Upon addition of 0.01% hydrogen peroxide and 10 nM o-phenylenediamine (final concentrations) in Tris buffer, the increase in absorbance was monitored for one minute at 450 nm. The presence of eosonophils in the uterus was taken as an indication of estrogenic activity of a compound. The maximal velocity of a fifteen second interval was determined over the initial, linear portion of the reaction curve.

Although EE₂ caused a decrease in serum cholesterol when orally administered at 0.1 mg/kg/day, it also exerted a stimulatory action on the uterus so that EE₂ uterine weight was substantially greater than the uterine weight of untreated ovariectomized test animals. This uterine response to estrogen is well recognized in the art. Representative compounds of the present invention reduced serum cholesterol compared to the ovariectomized control animals. Moreover, relative to EE₂, representative compounds of the present invention have a diminished effect on uterine weight.

Compared to estrogenic compounds known in the art, the benefit of serum cholesterol reduction without as adverse of an affect on uterine weight is rare and desirable.

Relative to EE₂, which caused a substantial, expected increase in eosinophil infiltration, the representative compounds of the present invention did not increase the eosinophil infiltration, and in most cases had a significantly diminished effect.

In addition to the above demonstrated benefits of these representative compounds of the present invention, especially when compared to estradiol, the compounds tested were not estrogen mimetic.

MCF-7 Proliferation Assay

The affinity of a representative sample of the compounds of the present invention for the estrogen receptors was tested in a MCF-7 receptor proliferation assay. MCF-7 breast adenocarcinoma cells (ATCC HTB 22) were maintained in MEM (minimal essential medium, phenol red-free, Sigma Chemical Co., St. Louis, Mo.) supplemented with 10% fetal bovine serum (FBS) (v/v), L-glutamine (2 mM), sodium pyruvate (1 mM), HEPES [N-(2-hydroxyethyl) piperazine-N'-2-ethanesulfonic acid 10 mM], non-essential amino acids and bovine insulin (1 mg/mL) (maintenance medium). Ten days prior to assay, MCF-7 cells were switched to maintenance medium supplemented with 10% dextran-coated charcoal stripped fetal bovine serum (DCC-FBS) assay medium) in place of 10% FBS to deplete internal stores of steroids. MCF-7 cells were removed from maintenance flasks using cell dissociation medium (Ca⁺² /Mg⁺² free HBSS (phenol red-free) supplemented with 10 mM HEPES and 2 mM EDTA). Cells were washed twice with assay medium and adjusted to 80,000 cells/mL. Approximately 100 mL (8,000 cells) were added to flat-bottom microculture wells (Costar 3596) and incubated at 37° C. in a 5% CO₂ humidified incubator for 48 hours to allow for cell adherence and equilibration after transfer. Serial dilutions of drugs or DMSO as a diluent control were prepared in assay medium and 50 mL transferred to triplicate microcultures followed by 50 mL assay medium for a final volume of 200 mL. After an additional 48 hours at 37° C. in a 5% CO₂ humidified incubator, microcultures were pulsed with tritiated thymidine (1 mCi/well) for four hours. Cultures were terminated by freezing at -70° C. for 4 hours followed by thawing and harvesting of microcultures using a Skatron Semiautomatic Cell Harvester. Samples were counted by liquid scintillation using a Wallac BetaPlace b-counter.

Relative to 17b-estradiol's known effects on the proliferation of MCF-7, the representative compounds of the present invention demonstrated significantly less stimulatory activity. In most cases an inhibitory effect was observed.

Bone Sparing: Five Week Ovariectomized Rat Model

Seventy-five day old female Sprague Dawley rats (weight range of 275 g-350 g) were obtained from Charles River Laboratories (Portage, MI). The animals were either bilaterally ovariectomized (OVX) or exposed to a Sham surgical procedure at Charles River Laboratories, and then shipped the day following surgery. Upon arrival, these rats were housed in metal hanging cases in groups of three or four animals per cage, and had ad libitum access to food (Teklad diet, TD 89222, 0.5% calcium, 0.4% phosphorous; Madison, WI) and water. Room temperature was maintained at 22.2° C.±1.7° C. with a minimum relative humidity of 40%. The photoperiod in the room was twelve hours light and twelve hours dark.

Test compound preparation was the same as that described in the Estrogenicty assay above. After a one day acclimation period (two days post-OVX), dosing with test compounds was initiated. Oral gavages 20% CDX, representative compound of the invention (0.01 to 10 mg/kg), or 17a-ethynylestradiol (100 mg/kg) were delivered daily for consecutive days. On the evening following the final dose, the animals were fasted. The next morning the animals were aneshetized with a mixture of Ketaset® and Rompun® (67 and 6.7 mg/kg respectively). The animals were asphyxiated with carbon dioxide and the left femur was removed from each animal, cleaned and frozen for subsequent X-ray evaluation.

Bone Assay

The distal end of the femur was X-rayed using a Norland NXR-1200 X-ray machine with a voltage of 47 kV and contrast at 4.5. Digitalized X-ray images were transferred to a computer station and image analysis of the X-ray scan was conducted. Quantitation was achieved by measuring the total number of pixels in a standard region of interest proximal to the growth plate, over a gray scale range of zero to 60.

When the above assay was run with the compound of Example 16 at a dose of 1 mg/kg, the X-ray evaluation resulted in 29.4% protection of the femur from bone loss.

For the majority of the methods of the present invention, compounds of formula I are administered continuously, from 1 to 3 times daily.

The specific dose of a compound administered according to this invention will, of course, be determined by the particular circumstances surrounding the case including, for example, the compound administered, the route of administration, the state of being of the patient, and the pathological condition being treated. A typical daily dose will contain a nontoxic dosage level of from about 5 mg to about 600 mg/day of a compound of the present invention. Preferred daily doses generally will be from about 15 mg to about 100 mg/day.

The compounds of this invention can be administered by a variety of routes including oral, rectal, transdermal, subucutaneus, intravenous, intramuscular, and intranasal, the selection of which will be decided by the attending physician. These compounds preferably are formulated prior to administration. Thus, another aspect of the present invention is a pharmaceutical formulation comprising an effective amount of a compound of formula I, or a pharmaceutical salt thereof, and a pharmaceutical carrier, diluent, or excipient. The total active ingredients in such formulations comprises from 0.1% to 99.9% by weight of the formulation.

Pharmaceutical formulations of the present invention can be prepared by procedures known in the art using well known and readily available ingredients. For example, the compounds of formula I can be formulated with common excipients, diluents, or carriers, and formed into tablets, capsules, suspensions, powders, and the like. Examples of excipients, diluents, and carriers that are suitable for such formulations include the following: fillers and extenders such as starch, sugars, mannitol, and silicic derivatives; binding agents such as carboxymethyl cellulose and other cellulose derivatives, alginates, gelatin, and polyvinyl-pyrrolidone; moisturizing agents such as glycerol; disintegrating agents such as calcium carbonate and sodium bicarbonate; agents for retarding dissolution such as paraffin; resorption accelerators such as quaternary ammonium compounds; surface active agents such as cetyl alcohol, glycerol monostearate; adsorptive carriers such as kaolin and bentonite; and lubricants such as talc, calcium and magnesium stearate, and solid polyethyline glycols.

The compounds also can be formulated as elixirs or solutions for convenient oral administration or as solutions appropriate for parenteral administration, for example, by intramuscular, subcutaneous, or intravenous routes. Additionally, the compounds are well suited for formulation as sustained-release dosage forms and the like. The formulations can be so constituted that they release the active ingredient only or preferably in a particular physiological location, possibly over a period of time. The coatings, envelopes, and protective matrices may be made, for example, from polymeric substances or waxes.

The following formulation examples are illustrative only and are not intended to limit the scope of the present invention.

Formulations

In the formulations which follow, "active ingredient" means a compound of formula I, or a salt thereof.

Hard gelatin capsules are prepared using the following:

Formulation 1 Gelatin Capsules

    ______________________________________                                         Ingredient        Quantity (mg/capsule)                                        ______________________________________                                         Active ingredient 0.1-1000                                                       Starch, NF 0-650                                                               Starch flowable powder 0-650                                                   Silicone fluid 350 centistokes 0-15                                          ______________________________________                                    

The formulation above may be changed in compliance with the reasonable variations provided.

A tablet formulation is prepared using the ingredients below:

Formulation 2 Tablets

    ______________________________________                                         Ingredient       Quantity (mg/tablet)                                          ______________________________________                                         Active ingredient                                                                               .sup. 2.5-1000                                                  Cellulose, microcrystalline 200-650                                            Silicon dioxide, fumed 10-650                                                  Stearate acid 5-15                                                           ______________________________________                                    

The components are blended and compressed to form tablets.

Alternatively, tablets each containing 2.5 mg-1000 mg of active ingredient are made up as follows:

Formulation 3 Tablets

    ______________________________________                                         Ingredient         Quantity (mg/tablet)                                        ______________________________________                                         Active ingredient  25-1000                                                       Starch 45                                                                      Cellulose, microcrystalline 35                                                 Polyvinylpyrrolidone 4                                                         (as 10% solution in water)                                                     Sodium carboxymethyl cellulose 4.5                                             Magnesium stearate 0.5                                                         Talc 1                                                                       ______________________________________                                    

The active ingredient, starch, and cellulose are passed through a No. 45 mesh U.S. sieve and mixed thoroughly. The solution of polyvinylpyrrolidone is mixed with the resultant powders which are then passed through a No. 14 mesh U.S. sieve. The granules so produced are dried at 50° C.-60° C. and passed through a No. 18 mesh U.S. sieve. The sodium carboxymethyl starch, magnesium stearate, and talc, previously passed through a No. 60 U.S. sieve, are then added to the granules which, after mixing, are compressed on a tablet machine to yield tablets.

Suspensions each containing 0.1 mg-1000 mg of medicament per 5 ml dose are made as follows:

Formulation 4 Suspensions

    ______________________________________                                         Ingredient         Quantity (mg/5 ml)                                          ______________________________________                                         Active ingredient    0.1-1000  mg                                                Sodium carboxymethyl cellulose 50 mg                                           Syrup 1.25 mg                                                                  Benzoic acid solution 0.10 mL                                                  Flavor q.v.                                                                    Color q.v.                                                                     Purified water to 5 mL                                                       ______________________________________                                    

The medicament is passed through a No. 45 mesh U.S. sieve and mixed with the sodium carboxymethyl cellulose and syrup to form a smooth paste. The benzoic acid solution, flavor, and color are diluted with some of the water and added, with stirring. Sufficient water is then added to produce the required volume.

An aerosol solution is prepared containing the following ingredients:

Formulation 5 Aerosol

    ______________________________________                                                              Quantity (% by                                              Ingredient weight)                                                           ______________________________________                                         Active ingredient    0.25                                                        Ethanol 25.75                                                                  Propellant 22 (Chlorodifluoromethane) 70.00                                  ______________________________________                                    

The active ingredient is mixed with ethanol and the mixture added to a portion of the propellant 22, cooled to 30° C., and transferred to a filling device. The required amount is then fed to a stainless steel container and diluted with the remaining propellant. The valve units are then fitted to the container.

Suppositories are prepared as follows: Formulation 6

    ______________________________________                                         Ingredient    Quantity (mg/suppository)                                        ______________________________________                                         Active ingredient                                                                            250                                                                Saturated fatty acid 2,000                                                     glycerides                                                                   ______________________________________                                    

The active ingredient is passed through a No. 60 mesh U.S. sieve and suspended in the saturated fatty acid glycerides previously melted using the minimal necessary heat. The mixture is then poured into a suppository mold of nominal 2 g capacity and allowed to cool.

An intravenous formulation is prepared as follows:

Formulation 7 Intravenous Solution

    ______________________________________                                         Ingredient      Quantity                                                       ______________________________________                                         Active ingredient     50     mg                                                  Isotonic saline 1,000 mL                                                     ______________________________________                                    

The solution of the above ingredients is intravenously administered to a patient at a rate of about 1 mL per minute.

Formulation 8 Combination Capsule I

    ______________________________________                                         Ingredient    Quantity (mg/capsule)                                            ______________________________________                                         Active ingredient                                                                            50                                                                 Premarin 1                                                                     Avicel pH 101 50                                                               Starch 1500 117.50                                                             Silicon Oil 2                                                                  Tween 80 0.5                                                                   Cab-O-Sil 0.25                                                               ______________________________________                                    

Formulation 9 Combination Capsule II

    ______________________________________                                         Ingredient    Quantity (mg/capsule)                                            ______________________________________                                         Active ingredient                                                                            50                                                                 Norethynodrel 5                                                                Avicel pH 101 82.50                                                            Starch 1500 90                                                                 Silicon Oil 2                                                                  Tween 80 0.50                                                                ______________________________________                                    

Formulation 10 Combination Tablet

    ______________________________________                                         Ingredient     Quantity (mg/capsule)                                           ______________________________________                                         Active ingredient                                                                             50                                                                Premarin 1                                                                     Corn Starch NF 50                                                              Povidone, K29-32 6                                                             Avicel pH 101 41.50                                                            Avicel pH 102 136.50                                                           Crospovidone XL10 2.50                                                         Magnesium Stearate 0.50                                                        Cab-O-Sil 0.50                                                               ______________________________________                                     

We claim:
 1. A compound of formula I: ##STR10## wherein: m is 0 or 1;R and R¹ are independently hydroxy, halo, or O--Pg; X is C═O, CH--OH, CH₂, O, or S; Pg is independently at each occurrence a hydroxy protecting group; R² is substituted C₅ -C₇ cycloalkyl or N-substituted -pyrrolidin-2-yl, -pyrrolidin-3-yl, -piperidin-2-yl, -piperidin-3-yl, -piperidin-4-yl, -hexamethyleneimin-3-yl, or -hexamethyleneimin-4-yl; where the N-substituent is C₁ -C₆ alkyl; where the substituent on the C₅ -C₇ cycloalkyl group is selected from the group consisting of pyrrolidin-1-yl, piperidin-1-yl, or hexamethyleneimin-1-yl; provided that if m is 1, then R² is substituted C₅ -C₇ cycloalkyl; or a pharmaceutical salt or solvate thereof.
 2. The compound according to claim 1 where X is C═O, m is 0, and R² is N-methyl, N-ethyl, or N-hexyl -piperidin-4-yl, -hexamethyleneimin-3-yl, or -hexamethyleneimin-4-yl.
 3. The compound according to claim 2 where R and R¹ are independently hydroxy or O--Pg, and Pg is independently at each occurrence methyl, ethyl, t-butyl, phenyl, or benzyl.
 4. The compound according to claim 3 where R and R¹ are both hydroxy.
 5. The compound according to claim 1 where X is C═O, m is 0, and R² is N-hexyl -piperidin-3-yl.
 6. The compound according to claim 5 where R and R¹ are independently hydroxy or O--Pg, and Pg is independently at each occurrence methyl, ethyl, t-butyl, phenyl, or benzyl.
 7. The compound according to claim 6 where R and R¹ are both hydroxy.
 8. The compound according to claim 1 where X is C═O, m is 0, and R² is substituted cyclohexyl where the cyclohexyl group is substituted with a piperidin-1-yl or pyrrolidin-1-yl group.
 9. The compound according to claim 8 where R and R¹ are independently hydroxy or O--Pg and Pg is independently at each occurrence methyl, ethyl, t-butyl, phenyl, or benzyl.
 10. The compound according to claim 9 where R and R¹ are both hydroxy.
 11. A pharmaceutical formulation comprising a compound of claim 1 and a pharmaceutical carrier, diluent, or excipient.
 12. The formulation according to claim 11 where X is C═O.
 13. A method for inhibiting a pathological condition associated with estrogen deficiency which comprises administering to a woman in need of such inhibition an effective amount of a compound of claim
 1. 14. The method according to claim 13 where X is C═O.
 15. The method according to either claim 13 or claim 14 where the pathological condition is osteoporosis.
 16. The method according to either claim 13 or claim 14 where the pathological condition is related to a cardiovascular disease.
 17. A method for inhibiting estrogen dependent cancer which comprises administering to a woman in need of such inhibition an effective amount of a compound of claim
 1. 18. The method according to claim 17 where the estrogen dependent cancer is breast cancer. 